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AB257121

Human CPS1 knockout HeLa cell lysate

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CPS1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 15 and Insertion of the selection cassette in exon 15.

View Alternative Names

CPSM_HUMAN, CPSase 1, CPSase I, Carbamoyl phosphate synthase, Carbamoyl phosphate synthase [ammonia] mitochondrial, Carbamoyl phosphate synthetase 1, Carbamoyl phosphate synthetase 1 mitochondrial, Carbamoyl-phosphate synthase [ammonia], Carbamoyl-phosphate synthetase I, MS738, mitochondrial

4 Images
Western blot - Human CPS1 knockout HeLa cell lysate (AB257121)
  • WB

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Western blot - Human CPS1 knockout HeLa cell lysate (AB257121)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : CPS1 knockout HeLa cell lysate (20 μg)

Lane 3 : Human liver tissue lysate (20 μg)

Lanes 1-3 : Merged signal (red and green). Green - ab155083 observed at 165 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab155083 Anti-CPS1 antibody [EPR7493-29] was shown to specifically react with CPS1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261809 (knockout cell lysate ab257121) was used. Wild-type and CPS1 knockout samples were subjected to SDS-PAGE. ab155083 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CPS1 antibody [EPR7493-29] (<a href='/en-us/products/primary-antibodies/cps1-antibody-epr7493-29-ab155083'>ab155083</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CPS1 knockout HeLa cell lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Predicted band size: 165 kDa

Observed band size: 165 kDa

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Western blot - Human CPS1 knockout HeLa cell lysate (AB257121)
  • WB

Unknown

Western blot - Human CPS1 knockout HeLa cell lysate (AB257121)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : CPS1 knockout HeLa cell lysate (20 μg)

Lane 3 : Human liver tissue lysate (20 μg)

Lanes 1-3 : Merged signal (red and green). Green - ab129076 observed at 165 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab129076 Anti-CPS1 antibody [EPR7493-3] was shown to specifically react with CPS1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261809 (knockout cell lysate ab257121) was used. Wild-type and CPS1 knockout samples were subjected to SDS-PAGE. ab129076 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CPS1 antibody [EPR7493-3] (<a href='/en-us/products/primary-antibodies/cps1-antibody-epr7493-3-ab129076'>ab129076</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CPS1 knockout HeLa cell lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Predicted band size: 165 kDa

Observed band size: 165 kDa

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Sanger Sequencing - Human CPS1 knockout HeLa cell lysate (AB257121)
  • Sanger seq

Unknown

Sanger Sequencing - Human CPS1 knockout HeLa cell lysate (AB257121)

Allele-2 : Insertion of the selection cassette in exon 15

Sanger Sequencing - Human CPS1 knockout HeLa cell lysate (AB257121)
  • Sanger seq

Unknown

Sanger Sequencing - Human CPS1 knockout HeLa cell lysate (AB257121)

Allele-1 : 1 bp insertion in exon 15

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 15 and Insertion of the selection cassette in exon 15.

Disease

Adenocarcinoma

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
CPS1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

CPS1 also known as carbamoyl-phosphate synthetase 1 is an important mitochondrial enzyme with a mass of around 165 kDa. It acts in the urea cycle by converting ammonia and bicarbonate into carbamoyl phosphate. CPS1 expression occurs in the liver and intestinal enterocytes which situates it in tissues responsible for metabolism and detoxification processes. This enzyme plays a critical role in the removal of excess nitrogen from the body.
Biological function summary

Carbamoyl-phosphate synthetase 1 functions as an initial step enzyme in the urea cycle which is a metabolic pathway for nitrogen disposal. It does not function alone as it forms part of multi-enzyme complexes to enhance efficiency in the cycle. These complexes facilitate the process of converting toxic ammonia into urea which the body can safely excrete. The proper function of CPS1 ensures that ammonia levels remain non-toxic.

Pathways

The urea cycle remains the chief pathway involving CPS1 collaborating with enzymes like ornithine transcarbamylase and argininosuccinate synthetase. This cycle intersects with the mitochondrial respiratory chain showcasing a relationship between CPS1 and cellular energy balance. Besides the urea cycle the enzyme also links to other metabolic functions like gluconeogenesis given its prerequisite for maintaining a stable nitrogen homeostasis.

CPS1 deficiencies can lead to urea cycle disorders like hyperammonemia characterized by elevated ammonia levels in blood. This may result in neurological damage if untreated. Another condition related to CPS-1 is pulmonary arterial hypertension where studies suggest mishandling of ammonia may alter blood pressure regulation. Genetic mutations affecting the CPS1 protein often underlie these conditions highlighting the importance of genetic screening in affected individuals.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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